Development of Methodology to Maintain Primary Cultures of Normal and Malignant Human Prostatic Epithelial Cells in Vivo

摘要

Our objective is to develop a realistic preclinical model of prostate cancer by developing methodology that supports the survival, growth and differentiation of primary cultures of prostate cells in mice. During year I, we focused on the method of implantation and the implantation site as the most critical elements in achieving this goal. In light of increasing evidence that stem cells are the only cells in cancers that have potential for sustained self-renewal, we now believe that primary cultures in fact will never be capable of forming tumors, regardless of the method of implantation, unless they contain stem cells. Analysis of the primary cultures established by our routine protocols suggested that stem cells were not present. Therefore, we devoted year two of this project to developing new methodology to establish prostate cancer stem cells in primary culture. Key steps included defining a protocol to isolate single, viable cells that retained cell surface antigens from fresh human cancer specimens, isolating distinct subpopulations of single cells expressing putative stem cell antigens, and determining the culture conditions required for single cells to attach and proliferate. These achievements now allow us to return to our original objective of defining optimal in vivo conditions, but with primary cultures that have the requisite stem cells necessary for tumor formation.